WO 95/10516, published Apr. 20, 1995 discloses tricyclic compounds useful for inhibiting farnesyl protein transferase.
In view of the current interest in inhibitors of farnesyl protein transferase, a welcome contribution to the art would be compounds useful for the inhibition of farnesyl protein transferase. Such a contribution is provided by this invention.
This invention provides compounds useful for the inhibition of farnesyl protein transferase (FPT). The compounds of this invention are represented by the formula: 
or a pharmaceutically acceptable salt or solvate thereof, wherein:
a represents N or NOxe2x88x92;
R1 and R3 are the same or different and each represents halo;
R2 and R4 are the same or different and each is selected from H and halo, provided that at least one of R2 and R4 is H;
T is a substituent selected from SO2R or 
Z is O or S;
n is zero or an integer from 1 to 6;
R is alkyl, aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, or N(R5)2;
R5 is H, alkyl, aryl, heteroaryl or cycloalkyl.
The compounds of this invention: (i) potently inhibit farnesyl protein transferase, but not geranylgeranyl protein transferase I, in vitro; (ii) block the phenotypic change induced by a form of transforming Ras which is a farnesyl acceptor but not by a form of transforming Ras engineered to be a geranylgeranyl acceptor; (iii) block intracellular processing of Ras which is a farnesyl acceptor but not of Ras engineered to be a geranylgeranyl acceptor; and (iv) block abnormal cell growth in culture induced by transforming Ras.
The compounds of this invention inhibit farnesyl protein transferase and the farnesylation of the oncogene protein Ras. Thus, this invention further provides a method of inhibiting farnesyl protein transferase, (e.g., ras farnesyl protein transferase) in mammals, especially humans, by the administration of an effective amount of the compounds of formula 1.0. The administration of the compounds of this invention to patients, to inhibit farnesyl protein transferase, is useful in the treatment of the cancers described below.
This invention provides a method for inhibiting or treating the abnormal growth of cells, including transformed cells, by administering an effective amount of a compound of this invention. Abnormal growth of cells refers to cell growth independent of normal regulatory mechanisms (e.g., loss of contact inhibition). This includes the abnormal growth of: (1) tumor cells (tumors) expressing an activated Ras oncogene; (2) tumor cells in which the Ras protein is activated as a result of oncogenic mutation in another gene; and (3) benign and malignant cells of other proliferative diseases in which aberrant Ras activation occurs.
This invention also provides a method for inhibiting or treating tumor growth by administering an effective amount of the tricyclic compounds, described herein, to a mammal (e.g., a human) in need of such treatment. In particular, this invention provides a method for inhibiting or treating the growth of tumors expressing an activated Ras oncogene by the administration of an effective amount of the compounds of formula 1.0. Examples of tumors which may be inhibited or treated include, but are not limited to, lung cancer (e.g., lung adenocarcinoma), pancreatic cancers (e.g., pancreatic carcinoma such as, for example, exocrine pancreatic carcinoma), colon cancers (e.g., colorectal carcinomas, such as, for example, colon adenocarcinoma and colon adenoma), myeloid leukemias (for example, acute myelogenous leukemia (AML)), thyroid follicular cancer, myelodysplastic syndrome (MDS), bladder carcinoma, epidermal carcinoma, breast cancer and prostate cancer.
It is believed that this invention also provides a method for inhibiting or treating proliferative diseases, both benign and malignant, wherein Ras proteins are aberrantly activated as a result of oncogenic mutation in other genesxe2x80x94i.e., the Ras gene itself is not activated by mutation to an oncogenic formxe2x80x94with said inhibition or treatment being accomplished by the administration of an effective amount of a compound of formula 1.0 to a mammal (e.g., a human) in need of such treatment. For example, the benign proliferative disorder neurofibromatosis, or tumors in which Ras is activated due to mutation or overexpression of tyrosine kinase oncogenes (e.g., neu, src, abl, lck, and fyn), may be inhibited or treated by the tricyclic compounds described herein.
The compounds of formula 1.0 useful in the methods of this invention inhibit or treat the abnormal growth of cells. Without wishing to be bound by theory, it is believed that these compounds may function through the inhibition of G-protein function, such as ras p21, by blocking G-protein isoprenylation, thus making them useful in the treatment of proliferative diseases such as tumor growth and cancer. Without wishing to be bound by theory, it is believed that these compounds inhibit ras farnesyl protein transferase, and thus show antiproliferative activity against ras transformed cells.
As used herein, the following terms are used as defined below unless otherwise indicated:
MH+xe2x80x94represents the molecular ion plus hydrogen of the molecule in the mass spectrum;
Et (or ET)xe2x80x94represents ethyl (C2H5);
alkylxe2x80x94represents straight and branched carbon chains that contain from one to twenty carbon atoms, preferably one to six carbon atoms;
haloxe2x80x94represents fluoro, chloro, bromo and iodo;
cycloalkylxe2x80x94represents saturated carbocyclic rings branched or unbranched of from 3 to 20 carbon atoms, preferably 3 to 7 carbon atoms;
heterocycloalkylxe2x80x94represents a saturated, branched or unbranched carbocylic ring containing from 3 to 15 carbon atoms, preferably from 4 to 6 carbon atoms, which carbocyclic ring is interrupted by 1 to 3 hetero groups selected from xe2x80x94Oxe2x80x94, xe2x80x94Sxe2x80x94 or xe2x80x94NR9xe2x80x94 (wherein R9 can be, for example, xe2x80x94C(O)N(R10)2, xe2x80x94CH2C(O)N(R10)2, xe2x80x94SO2R10, xe2x80x94SO2N(R10)2, xe2x80x94C(O)R11, xe2x80x94C(O)xe2x80x94Oxe2x80x94R11, alkyl, aryl, aralkyl, cycloalkyl, heterocycloalkyl or heteroaryl; each R10 independently represents H, alkyl, aryl, or aralkyl (e.g., benzyl); and R11 is alkyl, aryl, aralkyl, heteroaryl or heterocycloalkyl)xe2x80x94(suitable heterocycloalkyl groups include 2- or 3-tetrahydrofuranyl, 2- or 3-tetrahydrothienyl, 2-, 3- or 4-piperidinyl, 2- or 3-pyrrolidinyl, 2- or 3-piperizinyl, 2- or 4-dioxanyl, etc.xe2x80x94with preferred heterocycloalkyl groups being 2-, 3- or 4-piperidinyl substituted with R10 on the piperidinyl nitrogen);
aryl (including the aryl portion of aryloxy and aralkyl)xe2x80x94represents a carbocyclic group containing from 6 to 15 carbon atoms and having at least one aromatic ring (e.g., aryl is a phenyl ring), with all available substitutable carbon atoms of the carbocyclic group being intended as possible points of attachment, said carbocyclic group being optionally substituted (e.g., 1 to 3) with one or more of halo, alkyl, hydroxy, alkoxy, phenoxy, CF3, amino, alkylamino, dialkylamino, xe2x80x94COOR11 or xe2x80x94NO2 (wherein R11 is H, alkyl, aryl, heteroaryl or cycloalkyl); and
heteroarylxe2x80x94represents cyclic groups, optionally substituted with R3 and R4, having at least one heteroatom selected from O, S or N, said heteroatom interrupting a carbocyclic ring structure and having a sufficient number of delocalized pi electrons to provide aromatic character, with the aromatic heterocyclic groups preferably containing from 2 to 14 carbon atoms, e.g., triazolyl, 2-, 3- or 4-pyridyl or pyridyl N-oxide (optionally substituted with R11 as defined above), wherein pyridyl N-oxide can be represented as: 
The following solvents and reagents are referred to herein by the abbreviations indicated: ethanol (EtOH); methanol (MeOH); acetic acid (HOAc or AcOH); ethyl acetate (EtOAc); N,N-dimethyl-formamide (DMF); trifluoroacetic acid (TFA); trifluoro-acetic anhydride (TFAA); 1-hydroxybenzotriazole (HOBT); 1-(3-dimethylaminopropyl)-3-ethyl carbodiimide hydrochloride (DEC); diisobutylaluminum hydride(DIBAL); and 4-methylmorpholine (NMM).
The positions in the tricyclic ring system are: 
Those skilled in the art will also appreciate that the S and R stereochemistry for the C-11 position of the tricyclic ring is as follows: 
Preferred halo atoms for R1, R2, R3, and R4 in formula 1.0 are selected from: Br, Cl or I, with Br and Cl being preferred.
Compounds of formula 1.0 include compounds of the formula: 
wherein R1 and R3 are the same or different halo and a and T are as defined above. Preferably, for these dihalo compounds, R1 and R3 are independently selected from Br or Cl, and more preferably R1 is Br and R3 is Cl.
Compounds of formula 1.0 include compounds of formulas 1.1 and 1.2:
wherein R1, R3 and R4 in formula 1.1 are halo, and R1, R2 and R3 in formula 1.2 are halo. Compounds of formula 1.1 are preferred.
Preferably, in formula 1.1, R1 is Br, R3 is Cl, and R4 is halo. More preferably, in formula 1.1, R1 is Br, R3 is Cl, and R4 is Br.
Preferably, in formula 1.2, R1 is Br, R2 is halo, and R3 is Cl. More preferably, in formula 1.1, R1 is Br, R2 is Br, and R3 is Cl.
Also, preferably, for the compounds of this invention, substituent a in Ring I represents N.
T is preferably xe2x80x94SO2methyl or a group 
wherein R is a 3-pydinyl N-oxide, 4-pyridinyl N-oxide, 4-piperdinyl, 3-piperdinyl or 3-pyrrolidinyl group, wherein the 4-piperdinyl, 3-piperdinyl or 3-pyrrolidinyl groups may be substituted on the piperindinyl or pyrrolidinyl nitrogen with a group R9 which can be, for example, xe2x80x94C(O)N(R10)2, xe2x80x94CH2C(O)N(R10)2, xe2x80x94SO2R10, xe2x80x94SO2N(R10)2, xe2x80x94C(O)R11, xe2x80x94C(O)OR11, alkyl, aryl, aralkyl, cycloalkyl, heterocycloalkyl or heteroaryl; each R10 independently represents H, alkyl, aryl, or aralkyl (e.g., benzyl); and R11 is alkyl, aryl, aralkyl, heteroaryl or heterocycloalkyl.
Those skilled in the art will appreciate that compounds of formula 1.0 include compounds of formulas 1.3 and 1.4:
with compounds of 1.3 being preferred for compounds of formula 1.1, and with compounds of Formula 1.4 being preferred for compounds of formula 1.2.
Thus, compounds of the invention include compounds of the formulas: 
Certain compounds of the invention may exist in different isomeric (e.g., enantiomers and diastereoisomers) forms. The invention contemplates all such isomers both in pure form and in admixture, including racemic mixtures. Enol forms are also included.
Certain compounds of formula 1.0 will be acidic in nature, e.g. those compounds which possess a carboxyl or phenolic hydroxyl group. These compounds may form pharmaceutically acceptable salts. Examples of such salts may include sodium, potassium, calcium, aluminum, gold and silver salts. Also contemplated are salts formed with pharmaceutically acceptable amines such as ammonia, alkyl amines, hydroxyalkylamines, N-methylglucamine and the like.
Certain basic compounds of formula 1.0 also form pharmaceutically acceptable salts, e.g., acid addition salts. For example, the pyrido-nitrogen atoms may form salts with strong acid, while compounds having basic substituents such as amino groups also form salts with weaker acids. Examples of suitable acids for salt formation are hydrochloric, sulfuric, phosphoric, acetic, citric, oxalic, malonic, salicylic, malic, fumaric, succinic, ascorbic, maleic, methane-sulfonic and other mineral and carboxylic acids well known to those in the art. The salts are prepared by contacting the free base form with a sufficient amount of the desired acid to produce a salt in the conventional manner. The free base forms may be regenerated by treating the salt with a suitable dilute aqueous base solution such as dilute aqueous NaOH, potassium carbonate, ammonia and sodium bicarbonate. The free base forms differ from their respective salt forms somewhat in certain physical properties, such as solubility in polar solvents, but the acid and base salts are otherwise equivalent to their respective free base forms for purposes of the invention.
All such acid and base salts are intended to be pharmaceutically acceptable salts within the scope of the invention and all acid and base salts are considered equivalent to the free forms of the corresponding compounds for purposes of the invention.
Intermdiates useful in the preparation of the compounds of the invention may be prepared according to the procedures described in WO 95/10516 published Apr. 20, 1995, in WO 96/30363 published Oct. 3, 1996, in U.S. Pat. No. 5,151,423 and by the methods described below.
Compounds of the invention can be prepared according to the reaction: 
In the reaction, the carboxylic acid (14.0) is coupled to the tricyclic amine (13.0) using amide bond forming conditions well known to those skilled in the art. The substituents are as defined for Formula 1.0. For example, carbodiimide coupling methods (e.g., DEC) can be used. For example, the carboxylic acid (14.0) can be reacted with the tricyclic amine (13.0) using DEC/HOBT/NMM in DMF at about 25xc2x0 C. for a sufficient period of time, e.g., about 18 hours, to produce a compound of Formula 1.0.
When T is SO2R, X is halo, preferably, chloro. 
The tricyclic piperadine compound is dissolved in an appropriate solvent such as DMF of THF. A base is added such as triethylamine, and the appropriate alkylsulfonylchloride, prepared by methods known in the art, is added to the reaction mixture at 0xc2x0 C. to ambient temperature with stirring. After 1-24 hours, the reaction mixture is added to water and the product extracted with a suitable solvent such as ethylacetate. The crude reaction product can then be chromatographed on a silica gel column.
Alkylaminosulfonamido derivatives can be prepared similarly: 
wherein the R5 groups may be the same or different and each is as defined above. In this reaction, the tricyclic piperadine compound is dissolved in an appropriate solvent such as DMF of THF. A base is added such as triethylamine, and the appropriate alkylaminosulfonylchloride, prepared by methods known in the art, is added to the reaction mixture at 0xc2x0 C. to ambient temperature with stirring. After 1-24 hours, the reaction mixture is added to water and the product extracted with a suitable solvent such as ethylacetate. The crude reaction product can then be chromatographed on a silica gel column.
The carboxylic acids (14.0) and the sulfonates (14.2) are generally known in the art or can be prepared by methods well known in the literature.
Compounds of Formula 13.0 can be prepared from compounds of formula 13.0a: 
wherein R6 is H, alky, carboalkoxy or any other group that can be converted into a group T. The compounds of formula 13.0a are prepared by methods known in the art, for example, by methods disclosed in WO 95/10516, in WO 96/30363 published Oct. 3, 1996, in U.S. Pat. No. 5,151,423 and those described below.
The double bond in the compounds of formula 13.0a can be cleaved by oxidation, e.g., by the method in Preparative Example 3 below, to give the ketones of formula 15.0 below: 
Compounds of Formula 13.0a wherein the C-3 postion of the pyridine ring in the tricyclic structure is substituted by bromo (i.e., R1 is Br) can also be prepared by a procedure comprising the following steps:
(a) reacting an amide of the formula 
wherein R5a is hydrogen and R6a is C1-C6 alkyl, aryl or heteroaryl; R5a is C1-C6 alkyl, aryl or heteroaryl and R6a is hydrogen; R5a and R6a are independently selected from the group consisting of C1-C6 alkyl and aryl; or R5a and R6a, together with the nitrogen to which they are attached, form a ring comprising 4 to 6 carbon atoms or comprising 3 to 5 carbon atoms and one hetero moiety selected from the group consisting of xe2x80x94Oxe2x80x94 and xe2x80x94NR9axe2x80x94, wherein R9a is H, C1-C6 alkyl or phenyl;
with a compound of the formula 
wherein R2, R3, and R4 are as defined above and R7a is Cl or Br, in the presence of a strong base to obtain a compound of the formula 
(b) reacting a compound of step (a) with POCl3 to obtain a cyano compound of the formula 
(c) reacting the cyano compound with a piperidine derivative of the formula 
wherein L is halo selected from the group consisting of Cl and Br, to obtain a ketone of the formula below: 
(d)(i) cyclizing the ketone under acid conditions (e.g., aluminum chloride, triflic acid, or sulfuric acid) to obtain a compound of Formula 13.0a wherein R6 is methyl, which can be cleaved to give the compound of formula 15.0,
Methods for preparing compounds of Formula 13.0a disclosed in WO 95/10516, in WO 96/30363 published Oct. 3, 1996, in U.S. Pat. No. 5,151,423 and described below employ a tricyclic ketone intermediate. Such intermediates of the formula 
wherein R1, R2, R3 and R4 are as defined above, can be prepared by the following process comprising:
(a) reacting a compound of the formula 
(i) with an amine of the formula NHR5aR6a, wherein R5a and R6a are as defined in the process above; in the presence of a palladium catalyst and carbon monoxide to obtain an amide of the formula: 
(ii) with an alcohol of the formula R10aOH, wherein R10a is C1-C6 lower alkyl or C3-C6 cycloalkyl, in the presence of a palladium catalyst and carbon monoxide to obtain the ester of the formula 
followed by reacting the ester with an amine of formula NHR5aR6a to obtain the amide;
(b) reacting the amide with an iodo-substituted benzyl compound of the formula 
wherein R2, R3, R4 and R7a are as defined above, in the presence of a strong base to obtain a compound of the formula 
(c) cyclizing a compound of step (b) with a reagent of the formula R8aMgL, wherein R8a is C1-C8 alkyl, aryl or heteroaryl and L is Br or Cl, provided that prior to cyclization, compounds wherein R5a or R6a is hydrogen are reacted with a suitable N-protecting group.
Compounds of Formula 1.0 wherein substituent a is NO (Ring I) can be made from compounds of Formula 13.0a using procedures well known to those skilled in the art. For example, the compound of Formula 13.0a can be reacted with m-chloroperoxybenzoic acid in a suitable organic solvent, e.g., dichloromethane (usually anhydrous) or methylene chloride, at a suitable temperature, to produce a compound of Formula 13.0b 
which can then be cleaved to provide a compound of formula 15.0 above.
Generally, the organic solvent solution of Formula 13.0a is cooled to about 0xc2x0 C. before the m-chloroperoxybenzoic acid is added. The reaction is then allowed to warm to room temperature during the reaction period. The desired product can be recovered by standard separation means. For example, the reaction mixture can be washed with an aqueous solution of a suitable base, e.g., saturated sodium bicarbonate or NaOH (e.g., IN NaOH), and then dried over anhydrous magnesium sulfate. The solution containing the product can be concentrated in vacuo. The product can be purified by standard means, e.g., by chromatography using silica gel (e.g., flash column chromatography).
Alternatively, compounds of Formula 1.0, wherein substituent a is NO, can be made from compounds of Formula 1.0, wherein substituent a is N, by the m-chloroperoxybenzoic acid oxidation procedure described above.
Those skilled in the art will appreciate that it is preferable to avoid an excess of m-chloroperoxybenzoic acid when the oxidation reaction is carried out on the compounds of formula 13.0a. In these reactions an excess of m-chloroperoxybenzoic can cause oxidation of the C-11 double bond.
Compounds of formula 1.0 wherein Z is S can be prepared from compounds of formula 1.0 wherein Z is O by treatment with a suitable sulfur transfer reagent such as Lawsson""s reagent.
Compounds of the invention having asymmetric carbons (e.g., compounds of the invention wherein X is CH or N have an asymmetric carbon at the C-11 position of the the tricyclic ring) can be separated into enantiomers by techniques known in the art, e.g., by chiral salt resolution or by chiral HPLC.
Compounds useful in this invention are exemplified by the following examples, which should not be construed to limit the scope of the disclosure. 
3,10-dibromo-8-chloro-6,11-dihydro-11-one-5H-benzo[5,6]-cycloheptyl-[1,2-b]pyridine (2gm, 4.98 mmol) was dissolved in 20 ml of dry tetrahydrofuran under a dry nitrogen atmosphere. 5ml of a 1.5 molar solution of N-methyl-piperidine-4-magnesium chloride was added and the reaction stirred for 18 hours. The reaction mixture was washed with saturated ammonium chloride, dried over magnesium sulfate, filtered and evaporated to a brown oil which was chromatographed on silica gel using 2.5% methanol/methylene chloride as the eluent to obtain 2.11 gm, 85% of 3,10-dibromo-8-chloro-6,11-dihydro-11-(1-methyl-4-piperidinyl)-5H-benzo[5,6]cycloheptyl[1,2-b]pyridin-11-ol. FABMS (M+H)=501
Combine 25.86 g (55.9 mmol) of 4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic acid ethyl ester and 250 mL of concentrated H2SO4 at xe2x88x925xc2x0 C., then add 4.8 g (56.4 mmol) of NaNO3 and stir for 2 hours. Pour the mixture into 600 g of ice and basify with concentrated NH4OH (aqueous). Filter the mixture, wash with 300 mL of water, then extract with 500 mL of CH2Cl2. Wash the extract with 200 mL of water, dry over MgSO4, then filter and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 10% EtOAc/CH2Cl2) to give 24.4 g (86% yield) of the product. m.p.=165-167xc2x0 C., Mass Spec.: MH+=506 (CI). Elemental analysis: calculatedxe2x80x94C, 52.13; H, 4.17; N, 8.29; foundxe2x80x94C, 52.18; H, 4.51; N, 8.16.
Combine 20 g (40.5 mmol) of the product of Step A and 200 mL of concentrated H2SO4 at 20xc2x0 C., then cool the mixture to 0xc2x0 C. Add 7.12 g (24.89 mmol) of 1,3-dibromo-5,5-dimethyl-hydantoin to the mixture and stir for 3 hours at 20xc2x0 C. Cool to 0xc2x0 C., add an additional 1.0 g (3.5 mmol) of the dibromohydantoin and stir at 20xc2x0 C. for 2 hours. Pour the mixture into 400 g of ice, basify with concentrated NH4OH (aqueous) at 0xc2x0 C., and collect the resulting solid by filtration. Wash the solid with 300 mL of water, slurry in 200 mL of acetone and filter to provide 19.79 g (85.6% yield) of the product. m.p.=236-237xc2x0 C., Mass Spec.: MH+=584 (CI). Elemental analysis: calculatedxe2x80x94C, 45.11; H, 3.44; N, 7.17; foundxe2x80x94C, 44.95; H, 3.57; N, 7.16
Combine 25 g (447 mmol) of Fe filings, 10 g (90 mmol) of CaCl2 and a suspension of 20 g (34.19 mmol) of the product of Step B in 700 mL of 90:10 EtOH/water at 50xc2x0 C. Heat the mixture at reflux overnight, filter through Celite(copyright) and wash the filter cake with 2xc3x97200 mL of hot EtOH. Combine the filtrate and washes, and concentrate in vacuo to a residue. Extract the residue with 600 mL of CH2Cl2, wash with 300 mL of water and dry over MgSO4. Filter and concentrate in vacuo to a residue, then chromatograph (silica gel, 30% EtOAc/CH2Cl2) to give 11.4 g (60% yield) of the product. m.p.=211-212xc2x0 C., Mass Spec.: MH+=554 (CI). Elemental analysis: calculatedxe2x80x94C, 47.55; H, 3.99; N, 7.56; foundxe2x80x94C, 47.45; H, 4.31; N, 7.49.
Slowly add (in portions) 20 g (35.9 mmol) of the product of Step C to a solution of 8 g (116 mmol) of NaNO2 in 120 mL of concentrated HCl (aqueous) at xe2x88x9210xc2x0 C. Stir the resulting mixture at 0xc2x0 C. for 2 hours, then slowly add (dropwise) 150 mL (1.44 mole) of 50% H3PO2 at 0xc2x0 C. over a 1 hour period. Stir at 0xc2x0 C. for 3 hours, then pour into 600 g of ice and basify with concentrated NH4OH (aqueous). Extract with 2xc3x97300 mL of CH2Cl2, dry the extracts over MgSO4, then filter and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 25% EtOAc/hexanes) to give 13.67 g (70% yield) of the product. m.p.=163-165xc2x0 C., Mass Spec.: MH+=539 (CI). Elemental analysis: calculatedxe2x80x94C, 48.97; H, 4.05; N, 5.22; foundxe2x80x94C, 48.86; H, 3.91; N, 5.18.
Combine 6.8 g (12.59 mmol) of the product of Step D and 100 mL of concentrated HCl (aqueous) and stir at 85xc2x0 C. overnight. Cool the mixture, pour it into 300 g of ice and basify with concentrated NH4OH (aqueous). Extract with 2xc3x97300 mL of CH2Cl2, then dry the extracts over MgSO4. Filter, concentrate in vacuo to a residue, then chromatograph (silica gel, 10% MeOH/EtOAc+2% NH4OH (aqueous)) to give 5.4 g (92% yield) of the title compound. m.p.=172-174xc2x0 C., Mass Spec.: MH+=467 (FAB). Elemental analysis: calculatedxe2x80x94C, 48.69; H, 3.65; N, 5.97; foundxe2x80x94C, 48.83; H, 3.80; N, 5.97
Combine 16.6 g (0.03 mole) of the product of Preparative Example 3, Step D, with a 3:1 solution of CH3CN and water (212.65 mL CH3CN and 70.8 mL of water) and stir the resulting slurry overnight at room temperature. Add 32.833 g (0.153 mole) of NaIO4 and then 0.31 g (2.30 mmol) of RuO2 and stir at room temperature give 1.39 g (69% yield) of the product. (The addition of RuO is accompanied by an exothermic reaction and the temperature climbs from 20xc2x0 to 30xc2x0 C.) Stir the mixture for 1.3 hrs. (temperature returned to 25xc2x0 C. after about 30 min.), then filter to remove the solids and wash the solids with CH2Cl2. Concentrate the filtrate in vacuo to a residue and dissolve the residue in CH2Cl2. Filter to remove insoluble solids and wash the solids with CH2Cl2. Wash the filtrate with water, concentrate to a volume of about 200 mL and wash with bleach, then with water. Extract with 6 N HCl (aqueous). Cool the aqueous extract to 0xc2x0 C. and slowly add 50% NaOH (aqueous) to adjust to pH=4 while keeping the temperature  less than 30xc2x0 C. Extract twice with CH2Cl2, dry over MgSO4 and concentrate in vacuo to a residue. Slurry the residue in 20 mL of EtOH and cool to 0xc2x0 C. Collect the resulting solids by filtration and dry the solids in vacuo to give 7.95 g of the product. 1H NMR (CDCl3, 200 MHz): 8.7 (s, 1H); 7.85 (m, 6H); 7.5 (d, 2H); 3.45 (m, 2H); 3.15 (m, 2H). 
Combine 15 g (38.5 mmol) of 4-(8-chloro-3-bromo-5,6-dihydro-11H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-ylidene)-1-piperidine-1-carboxylic acid ethyl ester and 150 mL of concentrated H2SO4 at xe2x88x925xc2x0 C., then add 3.89 g (38.5 mmol) of KNO3 and stir for 4 hours. Pour the mixture into 3 L of ice and basify with 50% NaOH (aqueous). Extract with CH2Cl2, dry over MgSO4, then filter and concentrate in vacuo to a residue. Recrystallize the residue from acetone to give 6.69 g of the product. 1H NMR (CDCl3, 200 MHz): 8.5 (s, 1H); 7.75 (s, 1H); 7.6 (s, 1H); 7.35 (s, 1H); 4.15 (q, 2H); 3.8 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.8 (m, 2H); 2.6-2.2 (m, 4H); 1.25 (t, 3H). 
Combine 6.69 g (13.1 mmol) of the product of Step A and 100 mL of 85% EtOH/water, then add 0.66 g (5.9 mmol) of CaCl2 and 6.56 g (117.9 mmol) of Fe and heat the mixture at reflux overnight. Filter the hot reaction mixture through celite(copyright) and rinse the filter cake with hot EtOH. Concentrate the filtrate in vacuo to give 7.72 g of the product. Mass Spec.: MH+=478.0
Combine 7.70 g of the product of Step B and 35 mL of HOAc, then add 45 mL of a solution of Br2 in HOAc and stir the mixture at room temperature overnight. Add 300 mL of 1 N NaOH (aqueous), then 75 mL of 50% NaOH (aqueous) and extract with EtOAc. Dry the extract over MgSO4 and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 20%-30% EtOAc/hexane) to give 3.47 g of the product (along with another 1.28 g of partially purified product). Mass Spec.: MH+=555.9. 1H NMR (CDCl3, 300 MHz): 8.5 (s, 1H); 7.5 (s, 1H); 7.15 (s, 1H); 4.5 (s, 2H); 4.15 (m, 3H); 3.8 (br s, 2H); 3.4-3.1 (m, 4H); 9-2.75 (m, 1H); 2.7-2.5 (m, 2H); 2.4-2.2 (m, 2H); 1.25 (m, 3H). 
Combine 0.557 g (5.4 mmol) of t-butylnitrite and 3 mL of DMF, and heat the mixture at to 60xc2x0-70xc2x0 C. Slowly add (dropwise) a mixture of 2.00 g (3.6 mmol) of the product of Step C and 4 mL of DMF, then cool the mixture to room temperature. Add another 0.64 mL of t-butylnitrite at 40xc2x0 C. and reheat the mixture to 60xc2x0-70xc2x0 C. for 0.5 hrs. Cool to room temperature and pour the mixture into 150 mL of water. Extract with CH2Cl2, dry the extract over MgSO4 and concentrate in vacuo to a residue. Chromatograph the residue (silica gel, 10%-20% EtOAc/hexane) to give 0.74 g of the product. Mass Spec.: MH+=541.0. 1H NMR (CDCl3, 200 MHz): 8.52 (s, 1H); 7.5 (d, 2H); 7.2 (s, 1H); 4.15 (q, 2H); 3.9-3.7 (m, 2H); 3.5-3.1 (m, 4H); 3.0-2.5 (m, 2H); 2.4-2.2 (m, 2H); 2.1-1.9 (m, 2H); 1.26 (t, 3H). 
Combine 0.70 g (1.4 mmol) of the product of Step D and 8 mL of concentrated HCl (aqueous) and heat the mixture at reflux overnight. Add 30 mL of 1 N NaOH (aqueous), then 5 mL of 50% NaOH (aqueous) and extract with CH2Cl2. Dry the extract over MgSO4 and concentrate in vacuo to give 0.59 g of the title compound. Mass Spec.: M+=468.7. m.p.=123.9xc2x0-124.2xc2x0 C.
The title compound can be cleaved by the methodology of Preparative Example 3 to prepare the corresponding 11-ketone having 3,10-dibromo-8-chloro substituents. 
Combine 40.0 g (0.124 mole) of the starting ketone and 200 mL of H2SO4 and cool to 0xc2x0 C. Slowly add 13.78 g (0.136 mole) of KNO3 over a period of 1.5 hrs., then warm to room temperature and stir overnight. Work up the reaction using substantially the same procedure as described for Preparative Example 2, Step A. Chromatograph (silica gel, 20%, 30%, 40%, 50% EtOAc/hexane, then 100% EtOAc) to give 28 g of the 9-nitro product, along with a smaller quantity of the 7-nitro product and 19 g of a mixture of the 7-nitro and 9-nitro compounds. 
React 28 g (76.2 mmol) of the 9-nitro product of Step A, 400 mL of 85% EtOH/water, 3.8 g (34.3 mmol) of CaCl2 and 38.28 g (0.685 mole) of Fe using substantially the same procedure as described for Preparative Example 2, Step C, to give 24 g of the product 
Combine 13 g (38.5 mmol) of the product of Step B, 140 mL of HOAc and slowly add a solution of 2.95 mL (57.8 mmol) of Br2 in 10 mL of HOAc over a period of 20 min. Stir the reaction mixture at room temperature, then concentrate in vacuo to a residue. Add CH2Cl2 and water, then adjust to pH=8-9 with 50% NaOH (aqueous). Wash the organic phase with water, then brine and dry over Na2SO4. Concentrate in vacuo to give 11.3 g of the product. 
Cool 100 mL of concentrated HCl (aqueous) to 0xc2x0 C., then add 5.61 g (81.4 mmol) of NaNO2 and stir for 10 min. Slowly add (in portions) 11.3 g (27.1 mmol) of the product of Step C and stir the mixture at 0xc2x0-3xc2x0 C. for 2.25 hrs. Slowly add (dropwise) 180 mL of 50% H3PO2 (aqueous) and allow the mixture to stand at 0xc2x0 C. overnight. Slowly add (dropwise) 150 mL of 50% NaOH over 30 min., to adjust to pH=9, then extract with CH2Cl2. Wash the extract with water, then brine and dry over Na2SO4. Concentrate in vacuo to a residue and chromatograph (silica gel, 2% EtOAc/CH2Cl2) to give 8.6 g of the product.